Non-small cell lung cancer (NSCLC) accounts for 85% of lung cancer cases. Furthermore, overexpression of EGFR and its mutation are associated with the majority of NSCLCs and have been implicated in the process of malignant transformation by promoting cell proliferation, cell motility and cell survival. Recently, chemokine receptor CXCR4 and its ligand CXCL12 have been shown to play an important role in lung cancer progression and metastasis. In our preliminary data, we have shown that cannabinoids derived from plants, such as D9-tetrahydrocannabinol, inhibit epidermal growth factor receptor (EGFR)-mediated signaling and synthetic cannabinoids, such as JWH-133, inhibit CXCR4-mediated signaling. Cannabinoids have been shown to mediate their effects through cannabinoid receptors CB1 and CB2. Therefore, our central hypothesis is that synthetic cannabinoids that bind to CB1/CB2 receptors could be used as novel strategies to inhibit EGF/EGFR-mediated and CXCL12/CXCR4-mediated growth and metastasis in NSCLC. In this proposal, we will further define the inhibitory properties of synthetic cannabinoids against NSCLC growth and metastasis. To this end, we will use an innovative multidisciplinary approach by taking advantage of transgenic and knockout mouse model systems. In aim 1, we will analyze the expression of CB1 and CB2 receptors in NSCLC patient samples. We will also analyze the role of synthetic cannabinoids against EGF/EGFR and CXCL12/CXCR4-induced growth and migration of NSCLC cell lines. In aim 2, we will evaluate the specific potential of synthetic cannabinoids against lung cancer growth and metastasis in various animal models. We will first analyze the effect of synthetic cannabinoids on tumor growth and metastasis in vivo in SCID mouse model systems. Furthermore, we plan to use CB1 and CB2 knockout mice to specifically analyze the role of CB1/CB2 receptors in lung cancer progression and metastasis. In addition, we propose to analyze the effect of synthetic cannabinoids that bind to CB1 and CB2 receptors on a genetically engineered mouse model system that overexpresses mutant EGFR and enhances lung cancer growth in these mice. This model represents the most disease- relevant model of human lung cancer to determine the anti-tumorigenic effects of synthetic CB1/CB2 agonists. We expect that the use of different animal model systems will help us clarify the therapeutic potential of synthetic cannabinoids against NSCLC. Finally, in aim 3, we will delineate the synthetic cannabinoid-induced and CB1/CB2-mediated molecular mechanisms that inhibit EGFR and CXCR4- mediated growth and metastasis of NSCLC. This investigation on the role of synthetic cannabinoids and cannabinoid receptors CB1 and CB2 can open new therapeutic strategies toward the treatment of highly-fatal and chemo-resistant NSCLC, which is especially imperative considering the poor prognosis of NSCLC patients. The proposed research can yield important novel future clinical therapies that will significantly improve lung cancer mortality and promote lung health.